Hypoxia enhances anti-fibrotic properties of extracellular vesicles derived from hiPSCs via the miR302b-3p/TGFβ/SMAD2 axis

BMC Medicine

Research Article

Marcin Piejko¹  Kinga B. Stopa²  Michał Sarna³  Zbigniew Madeja⁴  Milena Paw⁴  Paweł E. Ferdek⁴  Dawid Wnuk⁴  Kinga Nit⁴  Ewa Zuba-Surma⁴  Sylwia Bobis-Wozowicz⁴  Jacek J. Litewka⁵  Agnieszka A. Kusiak⁵  Ruba Hammad⁶  Toni Cathomen⁶  Olga Barczyk-Woznicka⁷  Kinga Fic⁸ 

[1] 3Rd Department of General Surgery, Jagiellonian University - Medical College, Kraków, Poland;Doctoral School of Exact and Natural Sciences, Jagiellonian University, Kraków, Poland;Małopolska Centre of Biotechnology, Jagiellonian University, Kraków, Poland;Faculty of Biochemistry, Biophysics and Biotechnology, Department of Biophysics, Jagiellonian University, Kraków, Poland;Faculty of Biochemistry, Biophysics and Biotechnology, Department of Cell Biology, Jagiellonian University, Kraków, Poland;Faculty of Biochemistry, Biophysics and Biotechnology, Department of Cell Biology, Jagiellonian University, Kraków, Poland;Doctoral School of Exact and Natural Sciences, Jagiellonian University, Kraków, Poland;Freiburg iPS Core Facility, Institute for Transfusion Medicine and Gene Therapy, Medical Center, University of Freiburg, Freiburg, Germany;Center for Chronic Immunodeficiency (CCI), University of Freiburg, Freiburg, Germany;Institute of Zoology and Biomedical Research, Department of Cell Biology and Imaging, Jagiellonian University, Kraków, Poland;Małopolska Centre of Biotechnology, Jagiellonian University, Kraków, Poland;

关键词: Extracellular vesicles;    Induced pluripotent stem cells;    Hypoxia;    Low oxygen;    Heart fibrosis;    Therapy;   

DOI  :  10.1186/s12916-023-03117-w

 received in 2023-06-20, accepted in 2023-10-16,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundCardiac fibrosis is one of the top killers among fibrotic diseases and continues to be a global unaddressed health problem. The lack of effective treatment combined with the considerable socioeconomic burden highlights the urgent need for innovative therapeutic options. Here, we evaluated the anti-fibrotic properties of extracellular vesicles (EVs) derived from human induced pluripotent stem cells (hiPSCs) that were cultured under various oxygen concentrations.MethodsEVs were isolated from three hiPSC lines cultured under normoxia (21% O2; EV-N) or reduced oxygen concentration (hypoxia): 3% O2 (EV-H3) or 5% O2 (EV-H5). The anti-fibrotic activity of EVs was tested in an in vitro model of cardiac fibrosis, followed by a detailed investigation of the underlying molecular mechanisms. Sequencing of EV miRNAs combined with bioinformatics analysis was conducted and a selected miRNA was validated using a miRNA mimic and inhibitor. Finally, EVs were tested in a mouse model of angiotensin II-induced cardiac fibrosis.ResultsWe provide evidence that an oxygen concentration of 5% enhances the anti-fibrotic effects of hiPS-EVs. These EVs were more effective in reducing pro-fibrotic markers in activated human cardiac fibroblasts, when compared to EV-N or EV-H3. We show that EV-H5 act through the canonical TGFβ/SMAD pathway, primarily via miR-302b-3p, which is the most abundant miRNA in EV-H5. Our results show that EV-H5 not only target transcripts of several profibrotic genes, including SMAD2 and TGFBR2, but also reduce the stiffness of activated fibroblasts. In a mouse model of heart fibrosis, EV-H5 outperformed EV-N in suppressing the inflammatory response in the host and by attenuating collagen deposition and reducing pro-fibrotic markers in cardiac tissue.ConclusionsIn this work, we provide evidence of superior anti-fibrotic properties of EV-H5 over EV-N or EV-H3. Our study uncovers that fine regulation of oxygen concentration in the cellular environment may enhance the anti-fibrotic effects of hiPS-EVs, which has great potential to be applied for heart regeneration.Graphical Abstract

【 授权许可】

CC BY   
© The Author(s) 2023

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